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In Vitro antibacterial effects of antilipopolysaccharide DNA aptamer-C1qrs complexes

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Abstract

DNA aptamers were developed against lipopolysaccharide (LPS) from E. coli O111:B4 and shown to bind both LPS and E. coli by a colorimetric enzyme-based microplate assay. The polyclonal aptamers were coupled to human C1qrs protein either directly using a bifunctional linker or indirectly using biotinylated aptamers and a streptavidin-C1qrs complex. Both systems significantly reduced colony counts when applied to E. coli O111:B4 and K12 strains across a series of 10× dilutions of the bacteria in the presence of human serum; it was diluted 1: 103 in order to avoid significant bacterial lysis by the competing alternate pathway of complement activation. A number of candidate DNA aptamer sequences were cloned and sequenced from the anti-LPS aptamer library for future screening of antibacterial or “antibiotic” potential and to aid in eventual development of an alternative therapy for antibiotic-resistant bacterial infections.

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Abbreviations

1XBB:

single strength binding buffer

MB(s):

magnetic microbead(s)

Apt(s):

aptamer(s)

P b :

p value between treatment groups

CFU:

colony-forming units

PBS:

phosphate-buffered saline

ds:

double-stranded

PCR:

polymerase chain reaction

EGS:

ethane-1,2-diol bis(sulfosuccinimidyl-succinate)

RT:

room temperature

SELEX:

systematic evolution of ligands by exponential enrichment

GVB:

gelatin veronal buffer

HSCP(s):

human serum complement protein(s)

SMC:

sulfosuccinimidyl 4-(N-maleimido-methyl)cyclohexane-1-carboxylate

IgG, IgM:

immunoglobulins G, M

KDO:

3-deoxy-D-manno-octulosonic acid

ss:

single-stranded

LPS:

lipopolysaccharide

TSA:

tryptic soy agar

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  1. Operational Technologies Corporation, 4100 NW Loop 410, Suite 230, San Antonio, Texas, 78229, USA

    J. G. Bruno, M. P. Carrillo & T. Phillips

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  1. J. G. Bruno
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  2. M. P. Carrillo
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  3. T. Phillips
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Correspondence to J. G. Bruno.

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Bruno, J.G., Carrillo, M.P. & Phillips, T. In Vitro antibacterial effects of antilipopolysaccharide DNA aptamer-C1qrs complexes. Folia Microbiol 53, 295–302 (2008). https://doi.org/10.1007/s12223-008-0046-6

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  • DOI: https://doi.org/10.1007/s12223-008-0046-6

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